Aim To study the effects of a novel psychoactive drug,modafinil,on the GABAA receptor-induced current in rat hippocampus pyramidal neurons.Methods Whole-cell patch-clamp recordings were applied in cultured hippocampus pyramidal neurons.Results The GABA-activated currents were blocked by bicuculline,a competitive antagonist of GABAA receptor.Modafinil could partly inhibit IGABA.Pre-treating hippocampus neurons with glibenclamide for 30 min,modafinil-induced inhibition on IGABA was removed.Conclusion Modafinil is different from other psychostimulants.The protection of modafinil may be via modulating ATP sensitive potassium channel activation.

The teaching method, teaching process and teaching model of programming course were reformed in Beijing University of Traditional Chinese Medicine by combining task-driven and flipped classroom teaching con-cept with BB network platform to provide a free and open environment for students in order to train their capability of analyzing and solving problems and the skills of independent learning and life-long learning. Good results were achieved in two rounds of task-driven plus flipped classroom teaching, which are of significance for the teaching model and teaching structure reform of other courses.

Objective:To analyze the aberrant methylation of p16 gene in sera from primary liver cancer patients and to evaluate the clinical significance. Methods:A methylation-specific PCR was performed for the detection of promoter hypermethylation of p16 gene in blood DNA from 64 cases of PLC patients,and to anamyze the relationship between the aberrant methylation of p16 gene and the clinical pathological data . Results:76.6%(49/64)of the sera from 64 cases of PLC patients showed hypermethylation for p16 promoter,whereas no methylated p16 gene promoter were found in sera from liver benign diseases patients and normal control. Methylated p16 gene promoter in sera did not strongly correlated with HBsAg, AFP,stage ,metastasis and differentiation in PLC. Conclusion:Detection of the aberrant methylation of p16 gene in blood DNA from PLC patients might offer an effective means for the earlier auxillary diagnosis of the malignancy.

Objective: To analyze the aberrant methylation of death-asscciated protein kinase (DAPK) gene in blood from non-small cell lung cancer (NSCLC) patients and to evaluate its clinical significance. Methods: A methylation-specific PCR was performed for the detection of promoter hypermethylation of DAPK gene in blood DNA from 65 cases of NSCLC patients,and the relation of the aberrant methylation of DAPK gene and the clinical pathological data was analyzed. Results: 30.8% (20/65) of the blood from 65 cases of NSCLC patients showed hypermethylation in DAPK promoter, whereas no methylated DAPK gene promoter was found in blood from normal control or patients with benign lung diseases. DAPK gene promoter methylation in blood did not strongly correlate with the pathological class, stage, metastasis and differentiation of NSCLC. Conclusion: Detection of the aberrant methylation of DAPK gene in blood DNA from NSCLC patients might offer an effective means for the earlier auxiliary diagnosis of the malignancy.

Objective To explore the optimal dose-effect relationship of gene transfer according to the expression of pEGFP in tumor cells under different parameters mediated by microbubble contrast agent and ultrasound in vivo.Methods C57BL/6J mice were inoculated subcutaneously in the middle of the right flank with hepal-6 tumor cells.The mixture of SonoVue and enhanced green fluorescent protein(EGFP)plasmid was injected into the tail vein of each mouse, groups were randomly established according to different output intensity( 1 W/cm2, 2 W/cm2, 3 W/cm2 ), exposure time ( 1 min, 5 min, 10 min) and volume of SonoVue(30 μl,60 μl,90 μl), the expression of EGFP in tumor cells under different parameter was examined by flow cytometry and fluorescence microscopy.Results The expression of pEGFP in tumor cells was significantly higher with the increase of exposure time,output intensity and volume of SonoVue (1 min vs 5 min, P＜0.05; 1 W/cm2 vs 2 W/cm2, P＜0.05 ; 30/μl vs 60 μl, P＜0.05).However, the transfection efficiency of EGFP can not be enhanced continuously with the increasing exposure time,output intensity and volume of SonoVue (5 min vs 10 min, P＞0.05; 2 W/cm2 vs 3 W/cm2, P＞0.05; 60 μl vs 90 μl, P＞0.05).The highest transfection effieieney was achieved under intensities was 2 W/cm2 [(21.02±1.45)%]with 5 min[(23.22±1.91)%] exposed to ultrasound, and 60 /μl SonoVue [ (21.02±1.45) % ].Conclusions Gene transfeetion efficiency changded with the difference parameters.2 W/cm2,5 min and 60 μl SonoVue led to the optimum dose-effect relationship of gene transfer in vivo .

Objective To determine whether the cross-sectional area (CSA)of the calf measured with MRI could stage lower ex-tremity lymphedema (LEL)secondary to gynecological cancer treatments.Methods 148 patients were enrolled in this research.116 females with unilateral LEL and 32 without LEL after gynecological cancer treatments underwent calf MRI and water displacement. Total soft tissue CSA (T),muscle CSA (M)and subcutaneous tissue CSA (S)of affected calf,and difference of T (DT),M (DM) and S (DS)between calves were obtained on MRI at mid-calf level.Volume of affected calf and difference of volume (DV)between calves were obtained by water displacement.Statistical analysis was performed to determine feasibility of MRI measurements for ac-cessing LEL.Results There were close correlations between volume and T or S of affected calf,and between DV and DT or DS of calves.The correlations of stages of LEL with T and S of affected calf as well as DT and DS were stronger than the volume of affect-ed calf and DV (P< 0.01).Multivariate analysis showed more significant differences in T and S than in volume of affected calf,and in DS than in DV between LEL stages (P< 0.05).No difference was found in volume of affected calf and in DV between stage 0 andⅠ. For staging LEL,DS showed the most profound discrimination ability among all measurable parameters.Conclusion DS of calves could be the most reliable parameter recommended for staging and early diagnosis of LEL.

Objective To elucidate the molecular basis on the differences of infectivity in infected cells between Sindbis virus(SINV:YN87448 virus)and Sindbis like virus(SINLV:XJ-160 virus).Methods Compare the E(glycoprotein)gene sequence and secondary structure of YN87448 virus and XJ-160 virus by bioinformatics analysis.Analyze the contribution of E gene to the biological differences between SINV and SINLV by constructing recombinant virus.Results By bioinformatics analysis,YN87448 virus and XJ-160 virus have the same genomic structure,which has 11 717 nt and 11 626 nt respectively.There are 82 amino acid differences between E gene of these two viruses,and showed scattered distribution.The main peak is basically the same for the hydrophobic of the E gene protein,but in some region existing small differences.The recombinant virus which exchanged the E gene of XJ-160 virus with YN87448 virus totally showed the biological character of YN87448 virus,either in the showing time of CPE,plaque forming time and plaque diameter,or in expression of functional proteins.Conclusion E gene plays a major role in the differences of infectivity in infected cells between SINV and SINLV,this result provide the molecular biological evidences for elucidating the biological differences between SINV and SINLV.

Objective To investigate the diagnostic value of the real-time tissue elastography (RTE)and AST/PLT ratio in-dex (APRI)for the assessment of the early stage liver fibrosis in chronic liver disease patients.Methods Totally 90 patients with chronic liver disease were enrolled in the present study.The RTE score and the area percentage of the blue color (AREA,one of the elastic characteristics),the AST and PLT were recorded.Compared the results among different stages of liver fibrosis reported by biopsy considered as the gold standard for the assessment of liver fibrosis.The accuracy of the diagnosis of liver fibrosis by RTE scores,AREA and APRI were compared.Results RTE scores,AREA and APRI increased with the stage of liver fibrosis. Taking ≥S1 as the diagnosis standard of the early stage liver fibrosis,the area under the curve of ROC for RTE scores,AREA and APRI were 0.88,0.92,0.76 (P <0.05).Conclusion RTE is a new and promising sonography-based noninvasive method for the assessment of hepatic fibrosis in patients with chronic liver disease.Combined RTE with APRI have great clinical significance in di-agnosing the early stage liver fibrosis in chronic liver disease patients.

Forty-eight patients with subclinical Cushing's syndrome(SCS)were evaluated.Eleven of them underwent adrenalectomy(Group 1)and the other 37 cases did not(Group 2).Serum and urine corticosol, plasma ACTH and parameters related to metabolic syndrome(such as waist circumference,blood pressure,blood lipids and fasting plasma glucose)were measured.The data at diagnosis were compared with those during the survey.The results indicated that patients with SCS had a significantly high prevalence of metabolic syndrome.The symptoms and signs of metabolic syndrome could be improved after removing the tumor.Otherwise there is no improvement,some patients will even develop into overt Cushing's syndrome.